Compounds selectively inhibiting gamma 9 delta 2 T lymphocytes

ABSTRACT

The invention concerns compounds of formula CH 3 —R 1 —(CH 2 ) 2 —R 2  wherein: R 1  is selected among a tertiary alcohol; a 1,2-diol; a halohydrine; an epoxide; an alkene; an aldehyde or an α-hydroxyaldehyde; and R 2  is selected among a methylenediphosphonate; a difluoromethylenediphosphonate; or a monofluoromethylenediphosphonate. The invention also concerns the uses of said compounds as selective inhibitors of Tγ9δ2 lymphocytes, and their uses, in particular for therapeutic purposes.

[0001] The invention relates to compounds selectively inhibiting theTγ9δ2 lymphocytes carriers of receivers at variable regions Vγ9 and Vδ2.

[0002] The Tγδ lymphocytes of primates present in the peripheral blood(humans, monkeys) represent, in the healthy individual, conventionally 1to 5% of the lymphocytes of the blood and play a role in the immunesystem. It has been shown that they recognize their antigenic ligands bydirect interaction with the antigen, without presentation by moleculesof CMH of a presenting cell. The Tγ9δ2 lymphocytes (sometimes alsocalled Tγ2δ2 lymphocytes) are the Tγδ lymphocytes carrying TCR receiversat variable regions Vγ9 and Vδ2. They represent the majority of the Tγδlymphocytes of human blood.

[0003] When they are activated, the Tγδ lymphocytes exert a strongcytotoxic activity unrestrained by CMH, particularly effective to killvarious types of cells, particularly pathogenic cells. Nevertheless, themassive activation of the Tγδ lymphocytes accompanying sometimes thedevelopment of certain pathologies, can have or lead to a pathogeniccharacter. Such is the case in particular for the auto-immune maladiessuch as plaque sclerosis (Wucherpfennig K. et al “γδT cell receptorrepertoire in acute multiple scerosis lesion” 1992, PNAS 89, 4588) orthe Beh

et malady (Yamashita N. et al “Role of γδT lymphocytes in thedevelopment of Beh

et disease” Clinical Experimental, Immunology, 107(2), 241-247).

[0004] Such is the case moreover for a certain number of bacterialpathologies such as brucellosis, tularemia, salmonelloses, tuberculosis,ehrlichiosis, or parasitic pathologies such as malaria (malarialattack), visceral leishmaniosis, toxoplasmosis (for example Morita C. T.et al, “Direct presentation of non peptide prenyl pyrophosphate antigensto human gamma delta T cells”, 1996, Research in Immunology, Vol. 147, p347-353).

[0005] Various antigens of Tγ9δ2 lymphocytes have been described(WO-9520673, U.S. Pat. No. 5,639,653, “Natural and synthetic non peptideantigens recognized by human γδT cells”, Yoshimasa Tanaka et al, Nature,375, 1995, pp 155-158). Nevertheless, these natural antigens are notcompletely identified. Moreover, it is known that the mechanism ofactivation of the Tγ9δ2 lymphocytes by these antigens is particular,because it does not imply any known molecule of CMH (major complex ofhistocompatibility). But the nature of this mechanism remainsunexplained, such that the problem of adjusting inhibitors of Tγ9δ2lymphocytes remains unsolved.

[0006] WO-95/20673 also indicates that the principals having aphosphatase enzymatic activity (phosphohydrolase phosphoric monoesterand/or pyrophosphatase nucleotide and/or phosphohydrolase phosphoricdiester) such as the alkaline phosphatase, are adapted to inhibit theantigenic activity of natural origin, the so-called TUBag, from amycobacterial extract, vis-à-vis Tγ9δ2 lymphocytes. Nevertheless, thisinhibition takes place by cleaving the antigens and thus does not act onthe Tγ9δ2 lymphocytes themselves. Moreover, it is not specific and posesproblems of uncontrollable secondary effects to the extent that thebiological or physiological media themselves include numerousphosphorylated compounds and natural phosphatase enzymatic activities.

[0007] The invention thus seeks to provide compounds for selectiveinhibition of the Tγ9δ2 lymphocytic stimulation, which is to sayspecific immunosuppressive compounds for Tγ9δ2 lymphocytes.

[0008] The invention seeks more particularly to provide such compoundswhich will be compatible, on the one hand, with administration to aprimate and, on the other hand, with considerations of profitability forindustrial use (which must be produced in a simple manner, in largequantities, at an acceptable cost on an industrial scale).

[0009] Moreover, it is also desirable that the inhibition of the Tγ9δ2lymphocytes for the treatment of an excess of activation of the Tγ9δ2lymphocytes does not destroy definitively the immune system of thepatient or of the lymphocytic biological medium. Thus, the inventionalso seeks to provide compounds having an inhibitory activity which willbe not only selective with respect to Tγ9δ2 lymphocytes, but alsoreversible, such that the activity of the Tγ9δ2 lymphocytes mayultimately be restored.

[0010] The invention also seeks to provide new phosphorated compoundsand their process for production.

[0011] The invention also seeks to provide applications of the compoundsaccording to the invention for the selective and reversible inhibitionof the Tγ9δ2 lymphocytes. More particularly, the invention seeks toprovide applications for the compounds according to the invention fortherapeutic use, of the applications of the compounds according to theinvention for diagnosis, and applications of the compounds according tothe invention for the experimental study of Tγ9δ2 lymphocytes, theirantigens or specific immunosuppressive agents.

[0012] The invention seeks particularly to provide a treatment forpathologies implying an activation of the Tγ9δ2 lymphocytes, andparticularly selected from malaria (malarial attack), visceralleishmaniosis, toxoplasmosis, brucellosis, tularemia, salmonelloses,tuberculosis, ehrlichiosis, auto-immune maladies such as sclerosis byplaques or the Beh

et malady.

[0013] To do this, the invention relates to new compounds of theformula:

CH₃—R₁—CH₂)₂—R₂  (I)

[0014] in which R₁ is selected from the following functions:

[0015] in which CAT+ represents one or more organic or mineral cations(including the proton) identical or different, in the same compound,

[0016] except for 3-methyl-3-butene-1-yl-difluoromethylenediphosphonate,and 3-methyl-3-butene-1-yl-methylenediphosphonate.

[0017] The compounds according to formula (I) of the invention are thefollowing (IUPAC nomenclature):

[0018] R₁: tertiary alcohol function:

[0019] 3-methyl-3-butanol-1-yl-methylenediphosphonate;

[0020] 3-methyl-3-butanol-1-yl-monofluoromethylenediphosphonate;

[0021] 3-methyl-3-butanol-1-yl-difluoromethylenediphosphonate;

[0022] R₁: 1,2 diol function:

[0023] 3-methyl-3,4-butanediol-1-yl-methylenediphosphonate;

[0024] 3-methyl-3,4-butanediol-1-yl-monofluoromethylenediphosphonate;

[0025] 3-methyl-3,4-butanediol-1-yl-difluoromethylenediphosphonate.

[0026] R₁: halohydrin function wherein X=Cl, Br, I:

[0027] 3-(chloromethyl)-3-butanol-1-yl-methylenediphosphonate;

[0028] 3-(chloromethyl)-3-butanol-1-yl-monofluoromethylenediphosphonate;

[0029] 3-(chloromethyl)-3-butanol-1-yl-difluoromethylenediphosphonate;

[0030] 3-(bromomethyl)-3-butanol-1-yl-methylene-diphosphonate;

[0031] 3-(bromomethyl)-3-butanol-1-yl-monofluoromethylenediphosphonate;

[0032] 3-(bromomethyl)-3-butanol-1-yl-difluoromethylenediphosphonate;

[0033] 3-(iodomethyl)-3-butanol-1-yl-monofluoromethylenediphosphonate;

[0034] 3-(iodomethyl)-3-butanol-1-yl-methylenediphosphonate;

[0035] 3-(iodomethyl)-3-butanol-1-yl-difluoromethylenediphosphonate.

[0036] R₁: epoxyd function:

[0037] 3,4-epoxy-3-methyl-1-butyl-methylenediphosphonate;

[0038] 3,4-epoxy-3-methyl-1-butyl-monofluoromethylenediphosphonate;

[0039] 3,4-epoxy-3-methyl-1-butyl-difluoromethylenediphosphonate.

[0040] R₁: alkene function:

[0041] 3-methyl-3-butene-1-yl-methylenediphosphonate;

[0042] 3-methyl-3-butene-1-yl-monofluoromethylenediphosphonate;

[0043] 3-methyl-3-butene-1-yl-difluoromethylenediphosphonate.

[0044] R₁: aldehyde function (R₃=H)

[0045] 3-formyl-1-butyl-methylenediphosphonate;

[0046] 3-formyl-1-butyl-monofluoromethylenediphosphonate;

[0047] 3-formyl-1-butyl-difluoromethylenediphosphonate.

[0048] R₁: α-hydroxyaldehyde (R₃=OH):

[0049] 3-formyl-3-butanol-1-yl-methylenediphosphonate;

[0050] 3-formyl-3-butanol-1-yl-monofluoromethylenediphosphonate;

[0051] 3-formyl-3-butanol-1-yl-difluoromethylenediphosphonate.

[0052] The 3-methyl-3-butene-1-yl-difluoromethylenediphosphonate hasbeen described by “phosphorylation of isoprenoid alcohols” V. JoDavisson et al., J. Org. Chem. 1986, 51, 4775.

[0053] The invention moreover relates to compounds of formula (I) above(including 3-methyl-3-butene-1-yl-difluoromethylenediphosphonate) as totheir uses as agents for the selective inhibition of Tγ9δ2 lymphocytes.

[0054] The invention relates more particularly to the compounds offormula (I) above, as to their uses as agents for the inhibition ofselective phosphoantigenic activation of Tγ9δ2 lymphocytes by aphosphated antigen (phosphoantigen), such as a natural antigen (forexample the Tubag disclosed by Wo 95/20673), or artificial antigens suchas IPP (3-methyl-3-butene-1-yl-pyrophosphate), a phosphohalohydrincompound such as BrHPP (3-(bromomethyl)-3-butanol-1-yl-diphosphate) orIHPP (3-(iodomethyl)-3-butanol-1-yl-diphosphate), or a phosphoepoxidcompound such as EpoxPP (3,4 epoxy-3-methyl-1-butyl-diphosphate).

[0055] Although the real mechanism for the inhibition of Tγ9δ2lymphocytes by the compounds of the invention is not definitely setforth, the work of the inventors permits believing that such a selectiveinhibition of the Tγ9δ2 lymphocytes can be obtained by compounds whichsatisfy the three following conditions:

[0056] 1) having a molecule of topologic form corresponding to formula(I),

[0057] 2) having an R₁ function adapted to form a covalent bond by areaction of the nucleophile substitution or addition type, or theelectrophile addition in the presence of Tγ9δ2 lymphocytes,

[0058] 3) having a group structurally analogous to a pyrophosphate, butadapted to inhibit the enzymatic hydrolysis of the terminal phosphatenecessary to the activation of Tγ9δ2 lymphocytes.

[0059] Such a compound can thus have the property of occupying theantigenic recognition sites of the Vγ9 Vδ2 receptors thanks toconditions 1) and 2), but preventing the transduction of the activationsignal to the lymphocyte because the enzymatic hydrolysis of theterminal phosphate, which the inventors think would be necessary forthis transduction, is inhibited.

[0060] The function R₁ is selected so as to be compatible withconditions 1) and 2) above and to permit obtaining the compoundaccording to the invention. The CH₃—R₁—(CH₂)₂— group must thus be anantigenic ligand of the T Vγ9 Vδ2 receptor. It can be isopentenyl, ofcourse, which is an antigenic ligand. The inventors have shown moreoverthat the other groups CH₃—R₁—(CH₂)₂— of the formula (I) mentioned abovealso permit obtaining inhibitors of the Tγ9δ2 lymphocytes.

[0061] The group R₂ is selected from structural analogs ofpyrophosphates that are unhydrolizable or weakly hydrolizable. Suchanalogs of the pyrophosphates are known per se (cf. “ATP analogs” by R.G. Yount (1975) Adv. Enzymol. Vol. 43, p 1-56; “Synthesis of monofluoro-and difluoro-methylenephosphonate analogues of sn-glycerol-3-phosphateas substrates for glycerol-3-phosphate dehydrogenase and the X-Raystructure of the fluoromethylenephosphonate moiety” by J. Nieschalk etal. (1996) Tetrahedon vol. 52 pl65-176; “The difluoromethylenephosphatemoiety as a phosphate mimic: X ray structure of 2 amino-1,1-difluoroethylphosphonic acid” by R. D. Chambers et al. (1990) J. Chem. Soc.Chem. Commun. vol. 15, p 1053-1054).

[0062] A group R₂ should also be selected to be compatible with thesynthesis of the compound according to the invention.

[0063] The invention also relates to uses of the compounds according tothe invention as inhibitors for the Tγ9δ2 lymphocytes of primates,particularly as inhibitors of the proliferation and/or the cytotoxicactivity and/or the production of mediatory substances by the Tγ9δ2lymphocytes of the primates with TCR receptors comprising the variableregions Vγ9 and Vδ2.

[0064] The invention also relates to applications of the compoundsaccording to the invention for the treatment of cells sensitive to Tγ9δ2lymphocytes of primates, in a natural or artificial medium adapted tocontain Tγ9δ2 lymphocytes, in which said cells can be placed intocontact with these Tγ9δ2 lymphocytes, this medium being compatible withthe compounds according to the invention (which is to say it is notsusceptible to cause degradation at least under certain conditions oftreatment).

[0065] By “cell sensitive to Tγ9δ2 lymphocytes” is meant any cellsubject to the effective activity induced by Tγ9δ2 lymphocytes (cellulardeath, the invention permitting preventing destruction of the cells bylymphocytes); reception of salting out by the Tγ9δ2 lymphocytes (TNF-α,INF-γ . . . ); cellular proliferation induced by Tγ9δ2 lymphocytes.

[0066] The invention thus extends to a process for the selectiveinhibition of the Tγ9δ2 lymphocytes, particularly to a process forselective inhibition of the proliferation of Tγ9δ2 lymphocytes and/or ofthe cytotoxic activity of the Tγ9δ2 lymphocytes and/or the production ofmediatory substances by the Tγ9δ2 lymphocytes, in which these Tγ9δ2lymphocytes are placed in contact with at least one compound accordingto the invention in a medium containing Tγ9δ2 lymphocytes.

[0067] Preferably, and according to the invention, there is used atleast one compound according to the invention at a concentration in themedium which gives rise to a selective inhibition of the polyclonalproliferation of the Tγ9δ2 lymphocytes. This medium can be selected fromhuman blood, the blood of a non-human primate, extracts of human blood,and extracts of the blood of a non-human primate.

[0068] Preferably, and according to the invention, there is used aconcentration greater than the IC50 concentration of the compoundaccording to the invention, defined as that permitting reducing by 50%the intensity of the response of the Tγ9δ2 lymphocytes, according to theinduced cytotoxicity test, with a standard antigenic stimulant,particularly BrHPP at 80 nM.

[0069] Said medium can be extracorporeal, said inhibition processaccording to the invention being then an extracorporeal treatment, whichcan particularly take place in the laboratory, for example by thediagnosis or the study of the Tγ9δ2 lymphocytes or of their properties.For diagnosis, the inhibition of the Tγ9δ2 lymphocytes can serve toevaluate the condition of activation of the Tγ9δ2 lymphocytes removedfrom a patient, according to their behavior after placing them incontact with an inhibitory quantity of a compound according to theinvention.

[0070] Said medium can also be intracorporeal, the selective inhibitionof the Tγ9δ2 lymphocytes being then a therapeutic or diagnostic utility.

[0071] More particularly, said medium is the peripheral blood of aprimate. The invention thus includes in particular a process for theselective inhibition of Tγ9δ2 lymphocytes of the peripheral blood of aprimate—particularly human—in which there is administered a quantityadapted to inhibit the Tγ9δ2 lymphocytes, of at least one compoundaccording to the invention. There is thus administered at least onecompound according to the invention by any route—notably parenteral inthe peripheral blood—.

[0072] Said medium can also be a cellular site to be treated, and thereis administered at least one compound according to the inventiondirectly in contact with the cellular site to be treated (topicaladministration).

[0073] Thus, the invention includes applications of the compoundsaccording to the invention therapeutically for the curative orpreventive treatment of pathologies involving an activation of the Tγ9δ2lymphocytes of primates in a medium that can contain Tγ9δ2 lymphocytes.

[0074] The invention thus also relates to the compounds of the formula(I) for their use as active therapeutic substances in primates. Theinvention also relates to the use of the compounds according to formula(I), for their use in a therapeutic composition adapted to beadministered to a primate for the preventive or curative treatment of apathology involving the activation of Tγ9δ2 lymphocytes.

[0075] The invention relates in particular to therapeutic uses of thecompounds according to the invention for the treatment of pathologies ofprimates belonging to the group formed by parasitoses selected frommalaria (paludism), visceral leishmaniosis and toxoplasmosis;auto-immune maladies—particularly plaque scleroses and the Beh

et malady—involving an activation of the Tγ9δ2 lymphocytes; bacterialpathologies selected from brucellosis, tularemia, salmonelloses,tuberculosis, and ehrlichiosis. According to the invention, there isadministered a therapeutic composition adapted to release, in theperipheral blood and/or at a cellular site to be treated, a quantity ofat least one compound according to the invention adapted to inhibit theTγ9δ2 lymphocytes.

[0076] Thus, it has been shown generally in the prior art mentionedabove, that a composition having the property of inhibiting Tγ9δ2lymphocytes can be preferably used for the treatment of thesepathologies.

[0077] Conventionally, in all the texts, the terms “therapy” or“therapeutic” include not only the curative treatments or care, but alsothe preventive treatments (prophylaxis) such as vaccination. Thus, bypermitting selective inhibition of the Tγ9δ2 lymphocytes, the inventionpermits immunostimulation treatments that can preferably also serve asprophylaxis by preventing the development of Tγ9δ2 lymphocytes, as wellas curing by inhibiting Tγ9δ2 lymphocytes.

[0078] The invention thus also relates to a therapeutic or diagnosticcomposition comprising at least one compound according to the invention.More particularly, the invention relates to a therapeutic compoundcomprising a quantity suitable to be administered to aprimate—particularly in contact with the peripheral blood or by topicalroute—of at least one compound according to the invention—particularlyfor the preventive or curative treatment of the above-mentionedpathologies. A composition according to the invention can be animmunostimulatory composition, or a vaccine, the compounds according tothe invention being antigens selectively inhibiting the Tγ9δ2lymphocytes.

[0079] A therapeutic composition according to the invention can beprepared in galenic form adapted to be administered by any route,particularly by the parenteral route directly into the peripheral bloodof the primate, with at least one compound according to the invention ina quantity adapted to inhibit the Tγ9δ2 lymphocytes and one or severalsuitable excipients. Given the active concentration of the compoundsaccording to the invention (of the order of 10 to 1000 μM), such anadministration is to be envisaged without the risk of toxicity.

[0080] A therapeutic composition according to the invention can also beprepared in a suitable galenic form for its topical administration,directly in contact with the Tγ9δ2 lymphocytes.

[0081] The galenic form of a therapeutic composition according to theinvention is prepared according to the selected route of administration,by conventional techniques for galenic formulation. The quantity and theconcentration of the compound or compounds according to the invention,and the posology, are determined by reference to the knownchemotherapeutic treatments of the maladies to be treated, given thebioactivity of the compounds according to the invention relative to theTγ9δ2 lymphocytes, of the individual to be treated, and of the malady inquestion, and of the different biological effects.

[0082] Preferably, and according to the invention, there is administeredthe compound according to the invention in a quantity adapted to createin the peripheral blood of the patient a concentration greater than theIC50 concentration of the compound according to the invention as definedabove.

[0083] Preferably, and according to the invention, for a bioactivecompound at a concentration comprised between 1 μM and 1000 μM, there isadministered by any route a quantity of a compound or compoundsaccording to the invention comprised between 0.1 mg and 1 g—particularlybetween 1 mg and 100 mg—per kilogram of weight of the patient.

[0084] Moreover, it has been shown in vitro that the compounds accordingto the invention have no general toxicity. Moreover, it is known thatthe biochemical category of molecules to which the compounds accordingto the invention belong (phosphoesters) constitute a family of compoundscompatible with analogous and physiological biological media. Thecompounds according to the invention have thus no other toxic effectsthan those induced by their bioactivity on the Tγ9δ2 lymphocytes.

[0085] Moreover, the compounds according to the invention have asufficiently low molecular weight (particularly below 500) to becompatible with their elimination by renal or urinary route.

[0086] An example of formulation of an injectable therapeuticcomposition according to the invention for a primate of 1 kg is thefollowing:

[0087] 5 mg of sodium salt of3,4-epoxy-3-methyl-1-butyl-methylenediphosphonate (Epox-PCP) diluted in5 ml of sterile Ringer-Lactate buffer.

[0088] There is thus administered over 4 days: 1 dose per day of 5 mgfor 1 kg of animal, corresponding to a concentration in the circulatingblood of 50 mg/l, which can be greater than the IC50 concentration of 15μM for Epox-PCP (a concentration of 50 mg/l corresponding to about 160μM).

[0089] It is to be noted that most of the excipients or otherconventional acceptable pharmaceutical additives used, are chemicallycompatible with the compounds according to the invention.

[0090] A therapeutic composition according to the invention can alsopreferably comprise one or several other active principles, particularlyto provide a synergetic effect. In particular, a compound according tothe invention can serve as a vaccine adjuvant. The vaccine therapeuticcomposition according to the invention is thus comprised by a knownvaccine composition to which is added a quantity of compound accordingto the invention adapted to inhibit the Tγ9δ2 lymphocytes which will notbe able to exert their direct effective activity (for examplecytotoxic), nor regulatory of the Th-1 type (for example salting outinterferon and tumoral necrosis factor (TNF or “tumor necrosisfactor”)), and thereby promoting the lymphocyte B responses (for exampleproduction of antibodies).

[0091] The invention also extends to the use of at least one compoundaccording to the invention for the production of a therapeuticcomposition according to the invention. More particularly, the inventionbears on the use of at least one compound according to the invention forthe production of a therapeutic composition adapted for the preventiveor curative treatment of a pathology involving an activation of theTγ9δ2 lymphocytes of primates—particularly a pathology selected from thegroup mentioned above—. In this instance, the invention also extends tothe use of at least one compound according to the invention for theproduction of a therapeutic composition adapted to beadministered—particularly in contact with the peripheral blood or bytopical route—to a primate—notably human—for the preventive or curativetreatment of a pathology as mentioned above.

[0092] The invention also relates to a process for the production of acomposition—particularly a therapeutic composition—according to theinvention, having the property of selectively inhibiting Tγ9δ2lymphocytes, in which there is used at least one compound according tothe invention. The invention also relates to a process for theproduction of a therapeutic composition adapted for the preventive orcurative treatment of a pathology as mentioned above, in which there isused at least one compound according to the invention. The inventionbears in particular on a process for production of a therapeuticcomposition adapted to be administered—particularly in contact with theperipheral blood or by topical route, to a primate for the preventive orcurative treatment of a pathology such as mentioned above, in whichthere is used at least one compound according to the invention.

[0093] The compounds according to the invention can be preparedaccording to the reactions given hereafter according to the different R1and R2g groups.

[0094] In the reaction diagrams, PCP identifies themethylenediphosphonate group, PCHFP identifies themonofluoromethylenediphosphonate group, and PCF₂P identifies thedifluoromethylenediphosphonate group.

[0095] Reaction I:

[0096] For R₁: tertiary alcohol function, alkene, epoxyd, and R₂: PCP,PCHFP, PCF₂P:

[0097] in which Ts is tosyl, TsClis tosyl chloride, 4-DMAP is4-dimethylaminopyridine.

[0098] The tetrabutylammonium salts of the reagent R₂—H, used in aquantity at least equal to 2 molar equivalents, are, according to thegroup R₂ of the compound to be prepared:

[0099] for PCP: the tris(tetra-n-butylammonium)hydrogeno-methylene-diphosphonate prepared from methylene disphosphonicacid,

[0100] for PCF₂P: the tris(tetra-n-butylammonium)hydrogeno-difluoromethylene-diphosphonate prepared fromtetrakis(trimethylsilyl)-difluoromethylenedisphosphonate according tothe procedure described by V. Jo DAVISSON et al. J. Org. Chem, 51, p4768-4779, (1986),

[0101] for PCHFP: the tris(tetra-n-butylammonium)hydrogeno-monofluoromethylenediphosphonate prepared fromtetrakis(trimethylsilyl)-monofluoromethylenediphosphonate according tothe procedure described by J. NIESCHALK et al. (1996) Tetrahedron vol.52 pl65-176 and adapted according to V. Jo DAVISSON et al. J. Org.Chem., 51, p 4768-4779, (1986).

[0102] The alcohols (1) are commercially available products except thealcohol corresponding to the R1 epoxyd function which can be obtainedeasily (G. M. RUBOTTOM et al., Org. Synth. Coll. Vol 7, p 282 (1990),Wiley) by epoxydation of the alkene function as follows:

[0103] (1) wherein R₁: alkene function (1) wherein R₁: epoxyd function

[0104] Formula II:

[0105] For R₁: Halohydrine function (X=Cl, Br, I), and R₂: PCP, PCHFP,PCF₂P:

[0106] (3) wherein R₁: alkene function

[0107] Reaction III:

[0108] Variant for R₁: epoxyd function, and R₂: PCP, PCHFP, PCF₂P:

[0109] For R₁: 1,2-diol function, and R₂: PCP, PCHFP, PCF₂P:

[0110] (3) wherein R1: alkene function

[0111] in which KmnO₄ is potassium permanganate (in a quantity less thanor equal to 1 molar equivalent)

[0112] Reaction V:

[0113] For R₁: aldehyde function, and R₂: PCP, PCHFP, PCF₂P:

[0114] in which R₂—H is used in a quantity at least equal to 2 molarequivalents.

[0115] The compound (9) can be easily obtained in the form of an alcoholby the Grignard reaction between an alkenyl organomagnesium andformaldehyde or ethylene oxide, for example starting from1-chloro-2-methyl-3-butene.

[0116] Reaction VI:

[0117] For R₁: α-hydroxyaldehyde function, and R₂: PCP, PCHFP, PCF₂P:

[0118] in which PVPCC is Poly[vinyl(pyridinium chlorochromate)], asindicated by FRECHET J. M., WARNOCK J., and FARRALL J., J. Org. Chem,vol 43, N°13, p2618-21 (1978).

[0119] Other characteristics, objects, and advantages of the inventionwill become apparent from a reading of the examples which follow, givenby way of non-limiting example, and the accompanying drawings, in which:

[0120] FIGS. 1 to 6 are graphs representing the results obtained inExample 10,

[0121]FIG. 7 shows four graphs showing the results obtained in Example11,

[0122]FIG. 8 shows a graph showing the results obtained in Example 12,

[0123]FIG. 9 shows a graph showing the results obtained in Example 13,

[0124]FIGS. 10a, 10 b and 10 c show the results obtained in Example 14.

EXAMPLE 1 Production of 3-methyl-3-butene-1-yl-methylenediphosphonate(IPCP)

[0125] Preparation of 3-methyl-3-butene-1-yl-tosylate

[0126] Into a glass reactor provided for manipulation under an inertatmosphere and carefully dried, are introduced with magnetic agitation(2.32 mmoles-442 mg) of tosyl chloride and (2.55 mmoles-312 mg) of4-(N,N-dimethylamino)pyridine in 5 ml of anhydrous dichloromethane. Tothis mixture is added slowly with the help of a syringe and with meansof a septum (2.32 mmoles-200 mg) of isopentenol in solution in about 1ml of dichloromethane. The reaction is followed by chromatography onthin layer silica (silica gel 60 F-254 eluant: pentane/ethyl acetate85/15 v/v—Rf(R-Ots)=0.4 and Rf(TsCl)=0.5). After about 3 hours ofagitation under a nitrogen atmosphere, the reaction mixture is dilutedin a large volume of hexane (about 100 ml) which gives rise to theimmediate formation of a white precipitate. The mixture is then filteredand the filtrate concentrated by evacuation under reduced pressure. Thesolution is diluted with a little diethyl ether and again filtered.After evaporation of the solvent, there is obtained a yellowish oil. Theproduct is purified by chromatography on a preparative silica column(silica gel 60—eluant: pentane/ethyl acetate 85/15). (1.98 mmoles-475mg) of 3-methyl-3-butene-1-yl-tosylate (85% of yield as isolatedproduct) are thus obtained. The compound (colorless oil) is stored at+4° C. in an anhydrous medium.

[0127] Preparation of tris (tetra-n-butylammonium)hydrogeno-methylenediphosphonate:

[0128] There is prepared a solution containing (5.68 mmoles-1 g) ofmethylenediphosphonic acid in about 20 ml of deionized water. To thisacid solution (pH 1.0), there is added dropwise an aqueous solution oftetra-n-butylammonium hydroxide (BU₄NOH) at 40% by weight until there isobtained a pH value equal to 10.0. After lyophilization of the titratedsolution, there is obtained about 5 g of the salt oftetra-n-butylammonium (hygroscopic salt with an oily appearance) whichis dissolved in 10 ml of anhydrous acetonitrile. The saline solution isthen filtered and dried by successive evaporations of the solvent underreduced pressure. There is thus obtained a solution oftris(tetra-n-butylammonium) hydrogen-methylenediphosphonate with apurity equal to 97% (result deduced by analysis by ion chromatography—HPAEC). The volume is adjusted with the anhydrous acetonitrile so as toobtain a concentration of salt comprised between 0.5 and 1M. Thesolution is stored at −20° C. in anhydrous medium.

[0129] Preparation of 3-methyl-3-butene-1-yl-methylenediphosphonate(isopentenyl methylenediphosphonate):

[0130] In a carefully dried glass reactor, there is introduced under anitrogen atmosphere, 2.5 ml of a solution of tris(tetra-n-butylammonium)hydrogen-methylenediphosphonate of 0.7 M (1.75 mmoles) in anhydrousacetonitrile. The reactor is cooled by an ice bath and then there isadded with magnetic agitation and with the help of a syringe (0.70mmoles-168 mg) of 3-methyl-3-butene-1-yl-tosylate in solution in aminimum quantity of acetonitrile (0.5-1M). After introduction of thetosylate, the ice bath is withdrawn and then the reaction is continuedwith agitation at ambient temperature. The progress of the reaction isthen followed by ionic chromatography (HPAEC) on an IonPac® AS11 column.After about 3 hours, the solvent is evaporated under reduced pressureand the reaction medium redissolved in 3 ml of a mixture ofwater/2-propanol 98/2 (v/v). The solution is passed through a columncontaining (19 mequiv-4 g) of cationic resin DOWEX® 50-WX8-200 (NH₄ ⁺form) then eluted with 10 ml of the mixture of water (pH 9)/2-propanol98/2 (v/v). After lyophilization, there is recovered a white solidcontaining the raw product.

[0131] Purification:

[0132] Excess ammonium diphosphonate and a small proportion of inorganicsalts are separated from the reaction medium by co-precipitation in thepresence of ammonium hydrogencarbonate. The raw product obtained in thepreceding step is dissolved in 4 ml of ammonium hydrogencarbonate 0.1 Mwhich is transferred into a centrifugation tube of 25 ml. The solutionis then treated with 10 ml of a mixture of acetonitrile/2-propanol 1/1(v/v) by agitating the mixture vigorously (vortex) for several minutesuntil the formation of a precipitate. The tube is then centrifuged at2000 rpm at 10° C. for 5 minutes. The supernatant, in which areextracted the inorganic salts, is reserved at +4° C. The procedure isrepeated by redissolving the precipitate in 3 ml of ammoniumhydrogencarbonate 0.1 M to which are added 7 ml of theacetonitrile/2-propanol mixture. After elimination of the solvent fromthe combined supernatants in a rotative evaporator, there is obtained anoily liquid which is reserved at +4° C.

[0133] The ammonium tosylate is for the most part separated from thereaction mixture by extraction with the chloroform/methanol solvent 1/1(v/v). The oily liquid from the preceding step is dissolved in 4 ml ofdeionized water at pH 9 and treated with 1 ml of this solvent by aconventional extraction procedure repeated 3 times. Then there areeliminated from the aqueous phase the traces of solvent by evaporationunder reduced pressure at 30° C. The solution is stored at −20° C.

[0134] The product is ultimately purified as needed by ion exchangechromatography on cartridges of Sep-Pak Accell Plus QMA (Waters®) in anamount of 360 mg with 10 grams eluted successively with aqueoussolutions of ammonium hydrogencarbonate respectively of 20 mM, 40 mM,100 mM, then 200 mM followed b chromatography (HPAEC) of the elutedfractions. The fractions corresponding to the purified product arecombined and then lyophilized. For carrying out biological tests, theaqueous solutions of the product are sterilized by filtration on a 0.2μm filter and stored at −20° C. In the case of tests carried out invivo, the solutions are first passed over a cationic resin column DOWEX®50-WX8-200 (Na⁺ form) eluted with two volumes of the column of deionizedwater.

[0135] Analysis of the ammonium salt by mass spectrometry with so-called“electrospray” (negative mode) ionization:

[0136] ESI-MS: m/z=243 [M−H]⁻ pseudomolecular species

[0137] ESI-MS/MS of the [M−H]⁻ ion: m/z=225 (loss of H₂O); m/z=157(pyrophosphonate)

EXAMPLE 2 Production of3-(bromomethyl)-3-butanol-1-yl-methylenediphosphonate (BrHPCP)

[0138] 0.34 mmoles (100 mg) of3-methyl-3-butene-1-yl-methylenediphosphonate (ammonium salt) insolution in 2 ml of deionized water of neutral pH are treated under asuction hood with 1.9 ml of a saturated aqueous solution (0.18 M) ofbromene water (1 equivalent-0.34 mmoles of bromene). The bromene wateris added progressively and preferably to a cold solution of ammoniumsalt by acting by periodically agitating until the bromene water isdecolorized. In the case in which the bromene is added in slight excess(persistent yellow coloration), the solution is transferred into a glassflask and then placed for several minutes under reduced pressure(rotating evaporator) at a temperature of 30° C. until the colordisappears. The product,3-(bromomethyl)-3-butanol-1-yl-methylenediphosphonate is generatedquantitatively (0.33 mmoles-130 mg)—which result is deduced fromanalysis by ionic chromatography—HPAEC. The solution is then treated asin Example 1 for carrying out biological tests and stored at −20° C.

[0139] Analysis of the ammonium salt by mass spectrometry withionization, so-called “electrospray” (negative mode.):

[0140] ESI-MS: m/z=339,341 natural isotopes of bromene present in thepseudomolecular species [M−H]⁻

[0141] ESI-MS/MS of the [M−H]⁻ ion: m/z=259 (intramolecularrearrangement)

EXAMPLE 3 Production of3-(iodomethyl)-3-butanol-1-yl-methylenediphosphonate (1HPCP)

[0142] Preparation of iodized water:

[0143] A solution of iodized water of the order of 0.5 to 1 mM isprepared by prolonged sonication (about 15 minutes) of several iodinecrystals in a solution of deionized water, with filtration. For testsbearing on the largest quantities, more concentrated iodine solutionscan be obtained by adding a small proportion of alcohol to the initialaqueous solution. The iodized water is then titrated with sodiumthiosulfate with the use of starch as a color indicator.

[0144] Preparation of3-(iodomethyl)-3-butanol-1-yl-methylene-diphosphonate:

[0145] 5 μmoles (1 ml of a 5 mM solution) of3-methyl-3-butene-1-yl-methylenediphosphonate prepared according toExample 1 in the form of the ammonium salt in aqueous or hydroalcoholicmedium of neutral pH, are treated at ambient temperature by the additionof 1 equivalent of iodine in aqueous solution (5 ml iodized water at 1mM). The solution is held for 30 minutes at ambient temperature, then 30minutes at +4° C. carrying out vigorous periodical agitation. Afterdecoloration of the iodized water, the product3-(iodomethyl)-3-butanol-1-yl-methylenediphosphonate is generatedquantitatively. For carrying out biological tests, the solution is firstconcentrated by lyophilization and treated as in Example 1.

EXAMPLE 4 Production of3,4-epoxy-3-methyl-1-butyl-methylenediphosphonate (Epox PCP)

[0146] There is treated at ambient temperature, 1 ml of an aqueoussolution containing (2 mg-5.1 mmoles) of3-(bromomethyl)-3-butanol-1-yl-methylenediphosphonate (ammonium salt)prepared according to Example 2, with 0.5 ml of an ammoniac molarsolution. The solution is maintained under agitation for several minutesand then lyophilized to eliminate the ammonia. The dry residue obtainedafter lyophilization is redissolved in 1 ml of deionized water andpurified by ion exchange chromatography on cartridges of Sep-Pak AccellPlus QMA (Waters®) of 360 mg as described in Example 1.

[0147] Analysis of the ammonium salt by mass spectrometry with so-called“electrospray” (negative mode) ionization:

[0148] ESI-MS: m/z=259 [M−H]⁻ pseudomolecular species

[0149] ESI-MS/MS of the [M−H]⁻ ion: m/z=241 (loss of H₂O); m/z=157(pyrophosphonate)

EXAMPLE 5 Production of 3-methyl-3-butanol-1-yl-methylenediphosphonate(tButOHPCP)

[0150] According to a procedure analogous to that described in Example1, there is prepared in a first step 3-methyl-3-butanol-1-yl-tosylatefrom 3-methyl-1,3-butanediol. The3-methyl-3-butanol-1-yl-methylenediphosphonate is obtained by reacting0.5 mmole of tosylate and 1 mmole of tris(tetra-n-butylammonium)hydrogen-methylenediphosphonate at ambient temperature for 24 hours. Thepurification procedure is identical to that described in Example 1.

[0151] Analysis of the ammonium salt by mass spectrometry with so-called“electrospray” (negative mode) ionization:

[0152] ESI-MS: m/z=,261 [M−H]⁻ pseudomolecular species

[0153] ESI-MS/MS of the [M−H]⁻ ion: m/z=243 (loss of H₂O) m/z=157(pyrophosphonate)

EXAMPLE 6 Production of3-methyl-3,4-butanediol-1-yl-methylenediphosphonate (Diol PCP)

[0154] In glass flask, there is introduced 1 ml of an aqueous solutionof neutral pH of the ammonium salt of3-methyl-3-butene-1-yl-methylenediphosphonate (3.4 mmoles-1 mg)—preparedaccording to Example 1—to this solution are added several fractions, 680μl of a cold solution of potassium permanganate of 5 mM (1equivalent-3.4 μmoles) while agitating periodically the solution in acold hcamber (+4° C.). After about 40 minutes of reaction during which abrown precipitate of manganese dioxide forms, there are added severalmicroliters of a saturated aqueous solution of isopantenol. Themanganese dioxide is separated from the reaction mixture bycentrifugation and then filtration. The filtrate is purified by ionexchange chromatography on cartridges of Sep-Pak Accell Plus QMA(Waters®) of 360 mg as described in Example 1.

EXAMPLE 7 Production of3-methyl-3-butene-1-yl-difluoromethylenediphosphonate (IPCF₂P)

[0155] This product is prepared as described in Example 1, by reactingin anhydrous acetonitrile, 0.5 mmole of 3-methyl-3-butene-1-yl-tosylatewith (3 equivalents-1.5 mmoles) of the salt oftris(tetra-n-butylammonium) prepared according to the protocol describedby V. Jo Davisson et al. J. Org. Chem., 1986, 51 p 4768-4779.

[0156] Analysis of the ammonium salt by mass spectrometry with so-called“electrospray” (negative mode) ionization:

[0157] ESI-MS: m/z=279 [M−H]⁻ pseudomolecular species

[0158] ESI-MS/MS of the [M−H]⁻ ion: m/z=261 (loss of H₂O); m/z=193(pyrophosphonate)

EXAMPLE 8 Production of3-(bromomethyl)-3-butanol-1-yl-difluoromethylenediphosphonate (BrHPCF₂P)

[0159] This product is obtained by a reaction of3-methyl-3-butene-1-yl-difluoromethylenediphosphonate (preparedaccording to Example 7) with bromanated water by following the processdescribed in Example 2.

[0160] Analysis of the ammonium salt by mass spectrometry with so-called“electrospray” (negative mode) ionization:

[0161] ESI-MS: m/z=375,377 natural isotopes of bromene present in thepseudomolecular species [M−H]⁻

[0162] ESI-MS/MS of the [M−H]⁻ ion: m/z=295 (intramolecularrearrangement)

EXAMPLE 9 Production of3,4-epoxy-3-methyl-1-butyl-difluoromethylenediphosphonate (Epox PCF₂P)

[0163] This product is obtained by treatment in basic medium of3-(bromomethyl)-3-butanol-1-yl-difluoromethylenediphosphonate (preparedaccording to Example 8) by following the procedure described in Example4.

[0164] Analysis of the ammonium salt by mass spectrometry with so-called“electrospray” (negative mode) ionization:

[0165] ESI-MS: m/z=295 [M−H]⁻ pseudomolecular species

[0166] ESI-MS/MS of the [M−H]⁻ ion: m/z=277 (loss of water); m/z=193(difluoromethylenediphosphonate)

EXAMPLE 10 Measurement of the Cytotoxic Activity of a Tγ9δ2 CloneActivated by 80 nM of BrHPP, or Unactivated

[0167] The specific cytotoxic activity of a clone of Tγ9δ2 lymphocytes,measured according to the induced cytotoxicity test, is compared, thisactivity being stimulated with 80 nM of the antigen3-(bromomethyl)-3-butanol-1-yl-diphosphate (BrHPP) (small black dots inthe upper left of FIG. 1), and considered as the reference response(100%), relative to that of a culture of clones that are not stimulated(0%) (small white dots FIG. 1).

[0168] The curves of FIG. 1 show the percentage of residual response(induced cytotoxicity test) obtained in cultures stimulated by 80 nM ofBrHPP in the presence of different concentrations (on the abscissa) ofthe compounds according to the invention, namely BrHPCHFP (whitetriangles), IHPCP (black triangles), PCP Diol (black circles), PCP Epox(crossed gray squares), tButOHPCP (black squares), and IPCP (whitesquares), as obtained in the preceding examples.

[0169] It will be noted that the addition of increasing concentrationsof these compounds inhibits up to 100% the reference response.

[0170] The tests carried out as indicated above on different compoundsaccording to the invention permit defining their IC50 concentrations,expressed in micromoles in the following table, leading to theinhibition of 50% of the reference response of the lymphocytesstimulated by 80 nM of the compound BrHPP according to the inducedcytotoxicity test. Compound μM IPCP 700 tButOH PCP 1000 Epox PCP 30 BrHPCP 15 IH PCP 15 I PCF₂P 1000 Epox PCF₂P 300 BrH PCF₂P 150

[0171] Other similar tests have also been carried out withmonofluorinated analogous compounds (in which the group R₂ ismonofluoromethylenediphosphonate) BrHPCHFP and Epox PCHFP. Thesecompounds are bioactive (which is to say selectively inhibit the Tγ9δ2lymphocytes), with a bioactivity of 30 μM for BrHPCHFP and of 50 μM forEpox PCHFP, for a concentration of BrHPP equal to 150 μM.

[0172]FIGS. 2 and 3 are graphs similar to FIG. 1 obtained by replacingthe BrHPP antigen with the IPP antigen (isopentenylpyrophosphate) at 325μM or, respectively, at 162 μM. The compounds according to the inventionused were in these examples IPCP (black squares), BrHPCP (whitetriangles) and IHPCP (black triangles). As will be seen, the inhibitionby the compounds according to the invention does not depend on theantigen used to stimulate the Tγ9δ2 lymphocytes.

[0173] FIGS. 4 to 6 show the results obtained with these same threecompounds according to the invention but when using as the antigenstimulating the Tγ9δ2 lymphocytes, the compound BrHPP at a varyingconcentration, respectively, of 150 μM, 75 μM and 37 μM. As will beseen, the compounds according to the invention produce the inhibition oflymphocytes in all cases, but at concentrations which vary in the samesense as the concentrations of stimulation antigen used. Statedotherwise, the greater the concentration of stimulating antigen, thegreater must be the concentration of the compound according to theinvention necessary to inhibit the lymphocytes.

EXAMPLE 11 Measurement of the Inhibitory Activity and its ReversibleCharacter, by the Test of Induced Cytotoxicity and the Test of TNFSalting Out

[0174]FIG. 7 shows four graphs showing the inhibition by the compoundaccording to the invention BrHPCP and the restoration of the stimulatingantigen activity of BrHPP.

[0175] The two left graphs are obtained by stimulating the Tγ9δ2lymphocytes as in Example 12, by adding first 9 nM of BrHPP antigen intothe culture medium, then by adding increasing concentrations (on theabscissa) of the compound according to the invention, BrHPCP. The tworight hand graphs are obtained by incorporating first of all 60 μM ofthe compound according to the invention, BrHPCP, into the culture mediumin contact with the Tγ9δ2 lymphocytes, then by adding increasingconcentrations (on the abscissa) of the antigen compound stimulatingBrHPP. The values obtained are represented by black circles. The blackcircles give the values obtained in the absence of the initial compound(BrHPP on the left graphs, BrHPCP on the right graphs). The upper graphsgive the percentage of residual response in the cultures (inducedcytotoxicity test). The lower graphs give the concentration of TNFsalted out in pg/ml.

[0176] As will be seen, the compound according to the invention BrHPCPinhibits the stimulation by BrHPP, but this inhibition is reversible tothe extent to which, after inhibition by the compound according to theinvention BrHPCP, the stimulation is restored by adding BrHPP.

[0177] This reversible character of the inhibition of the Tγ9δ2lymphocytes by the compounds according to the invention is importantfrom the therapeutic point of view. Thus, following a treatment ofmassive activation of pathogenic character of the Tγ9δ2 lymphocytes,thanks to a compound according to the invention (for example during amalaria attack or on a tumor), the immune system of the patient is notnecessarily definitively degraded and afterward can be rapidly restored.

EXAMPLE 12 BrHPCP is not an Inhibitor of Tγ8δ3 Lymphocytes

[0178] A test of induced cytotoxicity is carried out as in Example 12,but with a clone of Tγ8γ3 lymphocyte stimulated by a conventionalantigen of these lymphocytes (black circles), and in the presence of thecompound according to the invention BrHPCP in increasing concentrationsin the culture medium (black squares in FIG. 8). As is seen in FIG. 8,the compound according to the invention does not inhibit the Tγ8γ3lymphocytes. It is thus a specific inhibitor for Tγ9δ2 lymphocytes.

EXAMPLE 13

[0179] In this example, there is carried out a test of inducedcytotoxicity on target cells P815 by a clone of Tγ9δ2 lymphocytesstimulated either by phytohemaglutinin A (PHA), which is a non-specificstimulant and is non-phosphated, for the Tγ9δ2 lymphocytes, at 70 ng/mland at 24 ng/ml, or by the antigen BrHPP at 80 nM. The stimulant is usedalone (white bars in FIG. 9) or in the presence of the inhibitorcompound according to the invention, BrHPCP, at 70 μM (black bars inFIG. 9).

[0180] As will be seen, the compound according to the invention does notinhibit the lymphocytes activated by the non-specific stimulant PHA. Itthus inhibits the Tγ9δ2 lymphocytes only if they have first beenstimulated in a specific manner by a phosphated antigen (phosphoantigen)such as BrHPP.

EXAMPLE 14

[0181] A million Tγ9δ2 lymphocytes are deposited in a well of 10 μl of amicrophysiometer (CYTOSENSOR® apparatus sold by MOLECULAR DEVICES, USA).Their speed of metabolism given by the apparatus is measured each 30seconds. There is added in the wells a composition comprising either theantigen BrHPP at 0, 2, 10 and 100 nM (FIG. 10a), or BrHPCP—at 2, 10, 100μM (white circles, squares and triangles in FIG. 10b), or the antigenIHPP (3-(iodomethyl)-3-butanol-1-yl-diphosphate) at 10 nM (black circlesin FIG. 10b), as a reference, or a controlled inactive composition(white circles in FIG. 10c), or BrHPCP at 50 μM alone (white circles inFIG. 10c), or IPP (isopentenylpyrophosphate) alone at 30 μM (blackcircles in FIG. 10c), or IPP at 30 μM and BrHPCP at 50 μM (whitetriangles in FIG. 10c).

[0182] The time of addition of the compositions is represented by thearrow in FIGS. 10a, 10 b, 10 c.

[0183] As will be seen, compared to the response detected upon additionof the antigens, the compounds according to the invention do not inducea response (FIG. 10b) and decrease the response to phosphoantigens (FIG.10c) of the Tγ9δ2 lymphocytes.

[0184] Continuing the experiment of FIG. 10c over a long period of timealso shows that the time during which the Tγ9δ2 lymphocytes areactivated is also decreased in the presence of compounds according tothe invention.

1. A compound having the following formula: H₃C—R₁—CH₂)₂—R₂  (I) whereinR₁ is selected from the following group consisting of:

 wherein R₃ is H or OH, wherein R₂ is selected from the group consistingof

and wherein Cat+ represents one or more organic or mineral cations,comprising the proton, identical or different, in the same compound,excepting 3-methyl-3-butene-1-yl-difluoromethylenediphosphonate, and3-methyl-3-butene-1-yl-methylenediphosphonate.
 2. The compositionaccording to claim 1, wherein said compound is combination with anexcipient or pharmaceutical additive.
 3. A method for selectivelyinhibiting Tγ9δ2 lymphocytes, comprising: contacting said Tγ9δ2lymphocytes with an effective amount of the compound according toclaim
 1. 4. A method for treating a patient with a pathology thatactivates Tγ9δ2 lymphocytes, comprising: administering to said patientin need thereof an effective amount of the compound according toclaim
 1. 5. A method for treating a primate with a pathology thatactivates Tγ9δ2 lymphocytes, comprising: administering to said primatein need thereof an effective amount of the compound according toclaim
 1. 6. A method for treating parasitosis in a primate, comprising:administering to said primate in need thereof an effective amount of thecompound according to claim
 1. 7. The method according to claim 6,wherein said parasitosis is selected from the group consisting ofmalaria, visceral leishmaniosis, toxoplasmosis.
 8. A method for treatingan autoimmune malady in a primate, comprising: administering to saidprimate in need thereof an effective amount of the compound according toclaim
 1. 9. The method according to claim 8, wherein said malady isbehcet malady.
 10. A method for selectively inhibiting Tγ9δ2 lymphocytesin an extracorporeal medium, comprising: contacting said Tγ9δ2lymphocytes in said extracorporeal medium with an effective amount ofthe compound according to claim
 1. 11. A method for inhibitingpolyclonal proliferation of Tγ9δ2 lymphocytes, comprising: contactingsaid Tγ9δ2 lymphocytes with an effective amount of the compoundaccording to claim 1.